I have tried to transfect 2 different cell lines with this plasmid but got no success. First of all, I transfect HEK 293 cell line with its transfection reagent and although I could see cell had fluorescense after transfection all of them die when I start the selection process with puromicin. In the case of CAL-62 cell line, fluorescence is lower than in the latter line but you can tell some cells have been transfected. However, at the selection process they do not survive. Trying a different transfection method with lipofectamine 2000 does not help either...Is there anybody who has use this plasmid? Did it work for you?
Thanks in advance,
Hi Teresa, I have same problem as you. I purchased double nickase plasmid with special target gene from Santa Cruz. I transfected Bes2B and WI-38 by Fugene HD (Lipo*2000 is toxic for my cells). During the selection process, the most cells are died. I used lowest dosage *0.5ur/mL" to select transfected cells. I am wondering if you have solution for your question, can you let me know what kind of method that you use?
Thank you so much.
Hi Wen-Chi Chen
I am sorry for the delay of the answer, I hadn't seen your question until now. I am not working on that any more, currently I am working in another laboratory..So, I am sorry for not being able to give you much information since I left all the protocols there...
I didn't achieve my objective, I couldn't be able to perform the transfection succesfully... Hope you had been more lucky...
Kind regards!
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